Production of a recombinant hybrid molecule of cholera toxin‐B‐subunit and proteolipid‐protein‐peptide for the treatment of experimental encephalomyelitis

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Abstract
Mucosal administration of experimental autoimmune encephalomyelitis (EAE)‐specific autoantigens can reduce the onset of disease. To examine whether cholera toxin‐B‐subunit (CTB)‐conjugated EAE‐specific T‐cell epitope can reduce development of the autoimmune disease in mice, we produced a recombinant hybrid molecule of CTB fusion protein linked with proteolipid‐protein (PLP)‐peptide139‐151(C140S) at levels up to 0.1 gram per liter culture media in Bacillus brevis as a secretion‐expression system. Amino acid sequencing and GM1‐receptor binding assay showed that this expression system produced a uniformed recombinant hybrid protein. EAE was induced in SJL/J mice by systemic administration with the PLP‐peptide. When nasally immunized 5 times with 70 μg rCTB PLP‐peptide hybrid protein, mice showed a significantly suppressed development of ongoing EAE and an inhibition of both the PLP‐peptide‐specific delayed‐type hypersensitivity (DTH) responses and leukocyte infiltration into the spinal cord. In contrast, all mice given the PLP‐peptide alone or the PLP‐peptide with the free form of CTB did not suppress the development of EAE and DTH responses. These results suggest that nasal treatment with the recombinant B. brevis‐derived hybrid protein of CTB and autoantigen peptide could prove useful in the control of multiple sclerosis. © 2001 John Wiley & Sons, Inc. Biotechnol Bioeng 74: 62–69, 2001.